Objective To investigate the role and mechanism of retinoblastoma protein-associated proteins 48 (RBBP4) in HIV-1 latency.Methods CEM-Bru cells latently infected with HIV-1 were stimulated with 25 ng/ml of tumor necrosis factor alpha (TNF-α) in combination with 10 ng/ml of interleukin-2 (IL-2).Chromatin immunoprecipitation (ChIP) was performed to detect the changes in RBBP4 and in histone deacetylases 1 and 2 (HDAC1/2) binding to long terminal repeat (LTR).Binding activities of HDAC1/2 and RNA polymerase Ⅱ (RNA Pol Ⅱ) to LTR and acetylated histone H3 at LTR region were detected by ChIP after partially interfering the expression of RBBP4 in CEM-Bru cells with electroporation.Initiating and elongated transcripts were measured by RT-PCR.Results The binding activities of RBBP4 and HDAC1/2 to LTR in HIV-1 latently infected cells were enhanced significantly as compared with those in TNF-α and IL-2 co-stimulated cells.Fewer RBBP4 and HDAC1/2 bound to LTR following the interference of RBBP4 expression, which was accompanied with enhanced histone acetylation and strengthened binding activity of RNA Pol Ⅱ to LTR.Moreover, more initiating transcripts were detected in HIV-1 latently infected cells after the RBBP4 expression was interfered by electroporation.Conclusion RBBP4 contributes to the maintenance of HIV-1 latency, in which HDAC1 and HDAC2 might be involved.

Objective To investigate the effect of retinoblastoma binding protein 4 (RBBP4)in Sp1 -mediated HIV long terminal repeat(LTR)transcription.Methods RBBP4 expression vector and Sp1 expression vector were respectively co-transfected into 293 T cells with HIV promoter pHIV-LTR-Luc or Sp1 site mutated pHIV-LTR-sp1 -mut by liposome transfection,and the transfected cells were examined by dual luciferase reporter assay system.The effect of RBBP4 on the binding of Sp1 to LTR was further studied by chromatin immunoprecipitation (ChIP)and electrophoretic mobility shift assay (EMSA).Results The relative firefly luciferase activity activated by Sp1 was decreased from 62.5 to 16 at the dose of 500 ng of RBBP4 expression vector (t =14.52,P <0.01 ).When the Sp1 binding sites were mutated,the effects of 100,300 or 500 ng of RBBP4 expression vector on the firefly luciferase activity of HIV LTR were not statistically significance (t =1 .897,2.357 and 3.162,all P <0.05).ChIP results showed that when the binding of RBBP4 on HIV LTR increased,the binding of Sp1 on HIV LTR increased significantly (t =11 .93,P <0.01 ),while the reduced binding of RBBP4 on HIV LTR significantly attenuated the binding of Sp1 onto LTR(t =11 .38,P <0.01 ).The effect of RBBP4 on the binding of Sp1 to DNA in ChIP assays was further verified by EMSA assays.Conclusion RBBP4 can inhibit the Sp1 -mediated HIV LTR transcription in 293 T cells.

OBJECTIVETo explore the feasibility of preparing novel gypenosides long-circulating liposomes with PEG grafted on beta-sitosterol (PEG-Sito).

METHODSuccinicanhydride was adopted to connect beta-sitosterol and PEG 2000. Sphingomyelin and PEG-Sito was used as material to prepare gypenosides long-circulating liposomes by using ethanol injection method. Encapsulation efficiency was determined by using protamine precipitation method. H-NMR was used to verify the synthesis of PEG-Sito, the novel gypenosides long-circulating liposomes were characterized by particle size, zeta potential and atomic force microscope.

RESULTThe synthesis of PEG-Sito was verified by 1H-NMR. Encapsulation efficiency of long-circulating liposomes prepared by ethanol injection method was 74.3%, particle size was 288.1 nm, zeta potential was -20.25 mV, the morphology were round observed by AFM.

CONCLUSIONThe novel gypenosides long-circulating liposomes prepared with PEG-Sito was feasible, it had a high encapsulation efficiency and good morphology.

Drug Compounding ; methods ; Feasibility Studies ; Gynostemma ; chemistry ; Liposomes ; blood ; chemistry ; Plant Extracts ; chemistry ; Polyethylene Glycols ; chemistry ; Reproducibility of Results ; Sitosterols ; chemistry ; Sphingomyelins ; chemistry

OBJECTIVE:To evaluate the effect of CD44 on drug resistance of breast cancer patients. METHODS:62 breast cancer patients were selected,and the expression of CD44 was detected by immunohistochemical method. Drug resistance was eval-uated by WHO criteria,and the expression of CD44 in breast tissue was observed after observation. RESULTS:There were 48 pa-tients sensitive to CD44 and 14 patients with drug resistance,with statistical significance(P<0.05). After chemotherapy,the ex-pression of CD44 in caner tissue was higher than before chemotherapy,that of patients with drug resistance was higher than pa-tients sensitive to CD44,with statistical significance(P<0.01). CONCLUSIONS:CD44 antigen is closely related to chemothera-py resistance of breast cancer,and high expression of CD44 can be used as an important indicator for breast cancer chemotherapy.

Objective To investigate the influences of mutation at precore and basic core promoter(BCP) region in hepatitis B virus(HBV) on the immune response of specific cytotoxic T lymphocytes(CTL) in patients with chronic hepatitis B(CHB).Methods The number of specific CTL in peripheral blood mononuclear(PBMC) of CHB patients were tested by cytokine flow cytome- try(CFC) and HBV core18-27 peptide.HBV precore and BCP fragments were directly sequenced. Results Twenty-one(38.9%) samples were HBV precore G1896A mutation.Twenty-six(48.1%) samples were BCP region 1762/1764 combined mutation.Thirteen(24.1%) stains were three sites mutated simultaneously.Stimulated with HBV core 18-27 in vitro,the specific CTL level was signifi- cantly higher in the patients with G1896A mutation and BCP region mutation [(0.41?0.09)%, (0.36?0.08)%,(0.48?0.08)%,respectively]than those without mutation[(0.11?0.06)%, P

Objective By the method of multiple polymerase chain reaction (PCR),we intend to amplify different regions (DFR) of Yersinia pestis DNA,and to establish a multiple DFR genotyping technique for detection of Yersinia pestis.Methods According to the product size of 23 DFRs and pMT plasmid,24 primers were optimized and combined,then multiple primers in one PCR reaction system were added,and positive template DNA was amplified.Meanwhile,200 wild strain DNAs were amplified by multiple PCR and normal PCR,to verify the coincidence rate of the two methods.Results Totally 24 target segments were amplified through the positive DNA template.Through different permutation and combination,24 primers were optimized and combined into 9 groups.Totally 200 wild strain DNAs were used for verification,the coincidence rate of multiple PCR and normal PCR was 100％.Conclusions Multiple PCR is applicable and feasible for DFR genotyping of Yersinia pestis.It is an efficient,economic and high accuracy experimental method for large quantities of Yersinia pestis DFR genotyping.

OBJECTIVETo investigate the electrophysiological effect of Xin' an granule (XAG) on ventricular muscle cell in ischemic rabbits.

METHODSA total of 48 rabbits were divided into the normal group and the ischemic group, and then subdivided into three groups, the control group, the high and low-dose XAG groups, 8 in each group. Rabbits in the low-dose XAG group and the high-dose XAG group were gastrogavaged XAG at the daily dose of 0. 85 g/kg and 3.40 g/kg, while the others in the control group were given the equal dosage of normal saline. All the rabbits were treated three times per day for successive 10 days. The rabbit model of ischemia was established by intravenous injected with 2. 5 U/kg posterior pituitary injection. Five minutes later, the monophasic action potential (MAP) and electrocardiogram (ECG) of each rabbit in the different groups were recorded and compared.

RESULTS(1) To normal rabbits, XAG could significantly shorten the action 50% and 90% potential duration (APD)50 and APD90 of ventricular muscle cell (P < 0.05 ), and high-dose of XAG could significantly increased the Vmax of MAP(P <0. 05). (2) While to ischemic rabbits, XAG could significantly prolong APD50 and APD90, and significantly increased the action potential amplitude (APA) and Vmax of MAP (P < 0. 05).

CONCLUSION(1) XAG can significantly shorten APD50 and APD90 of ventricular muscle cell, and high-dose XAG significantly increase the Vmax of MAP of normal rabbits. (2) XAG can delay and alleviate the manifestation characteristics of action potential of ventricular muscle cell during ischemia.

Action Potentials ; drug effects ; Animals ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Electrocardiography ; Heart Ventricles ; cytology ; drug effects ; Medicine, Chinese Traditional ; Myocardial Ischemia ; physiopathology ; Myocytes, Cardiac ; drug effects ; physiology ; Rabbits

OBJECTIVETo investigate the effect of four traditional Chinese medicines distributed along lung meridian, namely Ephedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex, on TLR2 and NF-κB expressions in mice with lung heat syndrome, in order to study the intervention effect of the four traditional Chinese medicines (TCMs) on the lung heat syndrome.

METHODOne hundred KM mice were randomly divided into the normal control group, the model group, the Ephedrae Hebra group, the Zingiberis Rhizoma group, the Scutellariae Radix group and the Mori Cortex group (20, 10 g x kg(-1)), nasally dripped with streptococcus pneumoniae to establish the mouse lung heat syndrome model, and then administered with different TCMs. The expressions of TLR2, NF-κB p65 proteins in lung tissues were analyzed by the immunohistochemical method. The expressions of TLR2, NF-κB p65 mRNA were measured by real time PCR.

RESULTCompared with the normal control group, the expressions of TLR2 and NF-κB p65 proteins in lung tissues in the model group were higher (P < 0.01), and the expressions of TLR2 and NF-κB p65 mRNA in lung tissues were up-regulated (P < 0.01 or P < 0.05). Compared with the model group, Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group and the Scutellariae Radix high dose group showed decreased expression of TLR2 protein (P < 0.05 or P < 0.01); Ephedrae Hebra high and low dose groups, the Zingiberis Rhizoma low dose group, Scutellariae Radix high and low dose groups and Mori Cortex high and low dose groups showed reduced expression of NF-κB p65 protein (P < 0.05 or P < 0.01). Ephedrae Hebra high and low dose groups, Zingiberis Rhizoma high and low dose groups, Scutellariae Radix high dose group and Mori Cortex high dose group showed down-regulated expression of TLR2 mRNA (P < 0.01 or P < 0.05).

CONCLUSIONEphedrae Hebra, Zingiberis Rhizoma, Scutellariae Radix and Mori Cortex can induce the TLR2/NF-κB inflammatory signal pathways by down-regulating the expressions of TLR2 and NF-κB p65 in protein and mRNA, so as to alleviate the lung tissue injury in mice with lung heat syndrome.

Animals ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; classification ; pharmacology ; Female ; Gene Expression ; drug effects ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; microbiology ; Male ; Medicine, Chinese Traditional ; methods ; Meridians ; Mice ; Phytotherapy ; methods ; Plants, Medicinal ; chemistry ; classification ; Pneumonia, Pneumococcal ; drug therapy ; genetics ; metabolism ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Scutellaria baicalensis ; chemistry ; Species Specificity ; Syndrome ; Toll-Like Receptor 2 ; genetics ; metabolism ; Transcription Factor RelA ; genetics ; metabolism

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Bone Morphogenetic Protein 4 ; metabolism ; Carrier Proteins ; pharmacology ; Liver ; metabolism ; Liver Regeneration ; Male ; Rats ; Rats, Wistar

To standardize the harvest ways of Dendrobium officinale and improve the quality and yield of D. officinale, a field experiment was carried out to study the effect of two kinds of harvest ways, which were keeping some of the axial shoot and harvesting all of the shoot by the end of the year. Then, the agronomic traits and yield were measured and the contents of polysaccharides and extractum were determined. The results showed that the harvest ways significantly affected the growth of D. officinale. Keeping some of the axial shoot could significantly improved the number of sprout, stem length, internode number and the internodal length, which also triggered increase the weight of fresh stems, leaves and the total of them and dry stems in per unit area, but it could not promote the stem diameter and the polysaccharide content in stems. Keeping some of the axial shoot moderately was conducive to the improvement of the production of medicinal materials in the process of harvesting by promoting the germination and growth of new buds, and to ensure the polysaccharide content by regulating the illumination and the density of cultivation.
Agriculture ; methods ; Dendrobium ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry ; growth & development ; Plant Stems ; chemistry ; growth & development